METHODS IN ENZYMOLOGY
1 INTRODUCTION A BRIEF HISTORY OF ENZYMOLOGY GOAL OF ENZYMOLOGY: THE ELUCIDATION OF MECHANISM THE EMERGENCE OF MECHANISM FROM DATA 2 KINETICS OF SINGLE-SUBSTRATE ENZYMATIC REACTIONS THE DEPENDENCE OF INITIAL VELOCITY ON SUBSTRATE CONCENTRATION AND THE REQUIREMENT FOR AN E : S COMPLEX DERIVATION OF THE RATE EQUATION FOR A SINGLE-SUBSTRATE REACTION WITH THE ASSUMPTION OF RAPID EQUILIBRIUM DERIVATION OF RATE EQUATIONS USING THE STEADY-STATE ASSUMPTION METHODS OF ENZYME ASSAY ENZYME KINETICS PRACTICUM: ASSAY DEVELOPMENT, EXPERIMENTAL DESIGN, DATA COLLECTION, AND DATA ANALYSIS 3 KINETICS OF SINGLE-SUBSTRATE ENZYMATIC REACTIONS: SPECIAL TOPICS TRANSITION STATE THEORY AND FREE ENERGY DIAGRAMS KINETIC CONSEQUENCES OF AN ENZYME : SUBSTRATE COMPLEX REACTIONS WITH MORE THAN ONE INTERMEDIARY COMPLEX DEVIATIONS FROM MICHAELIS–MENTEN KINETICS KINETICS OF ENZYMATIC ACTION ON SUBSTRATES WITH MULTIPLE REACTIVE CENTERS 4 ENZYME INHIBITION: THE PHENOMENON AND MECHANISM-INDEPENDENT ANALYSIS ENZYME INHIBITION: THE PHENOMENON ENZYME INHIBITION: THE FIRST QUANTITATIVE STEPS ENZYME-INHIBITOR SYSTEMS MISBEHAVING CASE STUDIES 5 KINETIC MECHANISM OF INHIBITION OF ONE-SUBSTRATE ENZYMATIC REACTIONS IMPORTANCE IN DRUG DISCOVERY THEORETICAL CONSIDERATIONS ANALYSIS OF INITIAL VELOCITY DATA FOR ENZYME INHIBITION INHIBITION OF ONE-SUBSTRATE, TWO-INTERMEDIATE REACTIONS INHIBITION BY DEPLETION OF SUBSTRATE 6 TIGHT-BINDING, SLOW-BINDING, AND IRREVERSIBLE INHIBITION TIGHT-BINDING INHIBITION SLOW-BINDING INHIBITION IRREVERSIBLE INHIBITION 7 KINETICS OF TWO-SUBSTRATE ENZYMATIC REACTIONS BASIC MECHANISMS CONCEPTUAL UNDERSTANDING OF SEQUENTIAL MECHANISMS DERIVATION OF RATE EQUATIONS FOR SEQUENTIAL MECHANISMS PING-PONG MECHANISMS DETERMINING THE KINETIC MECHANISM FOR TWO-SUBSTRATE REACTIONS A CONCEPTUAL UNDERSTANDING OF THE SHAPES OF SECONDARY PLOTS MISTAKEN IDENTITY: RAPID EQUILIBRIUM RANDOM VERSUS STEADY-STATE ORDERED 8 KINETIC MECHANISM OF INHIBITION OF TWO-SUBSTRATE ENZYMATIC REACTIONS MECHANISMS OF INHIBITION OF TWO-SUBSTRATE REACTIONS INHIBITION BY SUBSTRATE ANALOGS ANALYSIS OF SEQUENTIAL REACTIONS IN WHICH INHIBITOR BINDS TO ENZYME : PRODUCT COMPLEXES DRIVING SAR PROGRAMS FOR TWO-SUBSTRATE ENZYMATIC REACTIONS CONTENTS 9 ALLOSTERIC MODULATION OF ENZYME ACTIVITY MECHANISMS OF ENZYME MODULATION KINETICS OF ALLOSTERIC MODULATION MEANING OF ? AND ? CASE STUDIES: DEPENDENCE OF ALLOSTERIC MODULATION ON STRUCTURAL FEATURES OF THE SUBSTRATE 10 KINETICS-BASED PROBES OF MECHANISM 101 PH DEPENDENCE OF ENZYMATIC REACTIONS 102 TEMPERATURE DEPENDENCE OF ENZYMATIC REACTIONS 103 VISCOSITY DEPENDENCE OF ENZYMATIC REACTIONS 104 KINETIC ISOTOPE EFFECTS ON ENZYME-CATALYZED REACTIONS